相关实验视频
Updated: Feb 4, 2026

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Substrate Generation for Endonucleases of CRISPR/Cas Systems
Published on: September 8, 2012
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通过从RNA获取CRISPR间隔器进行转录记录
Florian Schmidt1, Mariia Y Cherepkova1, Randall J Platt2,3
1Department of Biosystems Science and Engineering, ETH Zurich, Basel, Switzerland.
Nature
|October 5, 2018
概括
这项研究引入了一种基于CRISPR的新方法,以记录细胞RNA活动作为DNA. 这项技术可以随着时间的推移跟踪复杂的细胞行为和对各种刺激的反应.
科学领域:
- 分子生物学
- 遗传学
- 生物技术
背景情况:
- 了解细胞行为需要随着时间的推移跟踪分子事件.
- 现有的分子记录技术在捕捉各种刺激方面存在局限性.
研究的目的:
- 开发一种记录细胞内RNA活动的新方法.
- 为了实现基于DNA的转录信息存储,用于细胞状态分析.
主要方法:
- 利用CRISPR间隔器获取将细胞内RNA转化为DNA.
- 在大肠杆菌中应用该方法来记录对定义和复杂刺激的反应.
- 开发了一种深度测序方法 (Record-seq) 用于转录历史重建.
主要成果:
- 在对RNA病毒,任意序列和氧化应激反应中展示了可量化的转录记录.
- 展示了使用转录记录来分类复杂细胞行为的能力.
- 鉴定了涉及不同细胞反应的特定基因.
结论:
- 通过CRISPR获取间隔器,可以实现高分辨率的转录记录.
- 记录-seq可以重建细胞转录历史.
- 这项技术在理解复杂的生物状态和疾病方面具有潜在的应用.
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Overview
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Tissue-specific transcription factors contribute to diverse cellular functions in mammals. For example, the gene for beta globin, a major component of hemoglobin, is present in all cells of the body. However, it is only expressed in red blood cells because the transcription factors that can bind to the promoter sequences of the beta globin gene are only expressed in these cells. Tissue-specific transcription factors also ensure that mutations in these factors may impair only the function of...
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The transcription elongation is regulated via pausing of RNA polymerase on several occasions during transcription. In bacteria, these halts are necessary because the transcription of DNA into mRNA is coupled to the translation of that mRNA...
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